•PLGA-PEG NSs with or without HPβCD were prepared for FB topical delivery.•NSs showed a 9-fold improvement in FB skin retention over the control solution.•HPβCD NSs exhibited the strongest inhibition ...of TPA-induced edema on mouse ears.•NSs showed no irritation on the rabbit skin.•NSs could be good candidates for FB topical delivery with minimal side effects.
In this study, flurbiprofen (FB) loaded poly(d,l-lactide-co-glycolide) (PLGA) and PLGA with poly(ethylene glycol) (PLGA-PEG) nanospheres (NSs) with and without hydroxypropyl-β-cyclodextrin (HPβCD) were developed as skin controlled delivery systems. X-ray diffraction was used to determine the physical state of the entrapped drug. Results showed that the drug in PLGA NSs was present in the form of a molecular dispersion (dissolved state) in the polymers, whereas in PLGA-PEG NSs, the drug was present in both molecular dispersion and crystalline forms. Furthermore, HPβCD provided solubilization of the free FB present on the surface of the PLGA-PEG NSs and a complete amorphosization of the drug was obtained. Optical analyses using TurbiscanLab® demonstrated that HPβCD provided an efficient steric stability of the NSs, preventing particle aggregation. The ex vivo permeation profiles of the NSs and conventional FB solution were evaluated using human skin. Results demonstrated that only PLGA-PEG NSs showed slight permeation improvement. However, after 24h, the FB retained in the skin was about 9-fold higher with NSs compared with the control solution, attributed to the reservoir effect of NSs acting as a depot, sustaining the drug and limiting its systemic absorption. In vivo performance of NSs was evaluated by assessing anti-inflammatory efficacy in TPA-induced mouse ear edema. Topically applied NSs significantly decreased in vivo inflammation compared to the control solution and the anti-inflammatory efficacy of HPβCD NSs was stronger than NSs without HPβCD. In vivo skin irritation evaluated by the in vivo Draize test showed no irritation of the formulations tested.
Effective treatment for hepatitis C virus (HCV) in patients coinfected with human immunodeficiency virus type 1 (HIV-1) remains an unmet medical need.
We conducted a multicenter, single-group, ...open-label study involving patients coinfected with HIV-1 and genotype 1 or 4 HCV receiving an antiretroviral regimen of tenofovir and emtricitabine with efavirenz, rilpivirine, or raltegravir. All patients received ledipasvir, an NS5A inhibitor, and sofosbuvir, a nucleotide polymerase inhibitor, as a single fixed-dose combination for 12 weeks. The primary end point was a sustained virologic response at 12 weeks after the end of therapy.
Of the 335 patients enrolled, 34% were black, 55% had been previously treated for HCV, and 20% had cirrhosis. Overall, 322 patients (96%) had a sustained virologic response at 12 weeks after the end of therapy (95% confidence interval CI, 93 to 98), including rates of 96% (95% CI, 93 to 98) in patients with HCV genotype 1a, 96% (95% CI, 89 to 99) in those with HCV genotype 1b, and 100% (95% CI, 63 to 100) in those with HCV genotype 4. Rates of sustained virologic response were similar regardless of previous treatment or the presence of cirrhosis. Of the 13 patients who did not have a sustained virologic response, 10 had a relapse after the end of treatment. No patient had confirmed HIV-1 virologic rebound. The most common adverse events were headache (25%), fatigue (21%), and diarrhea (11%). No patient discontinued treatment because of adverse events.
Ledipasvir and sofosbuvir for 12 weeks provided high rates of sustained virologic response in patients coinfected with HIV-1 and HCV genotype 1 or 4. (Funded by Gilead Sciences; ION-4 ClinicalTrials.gov number, NCT02073656.).
Triatomines (Hemiptera: Reduviidae) are the insect vectors of Trypanosoma cruzi, the causative agent of Chagas disease. The gut bacterial communities affect the development of T. cruzi inside the ...vector, making the characterization of its composition important in the understanding of infection development. We collected 54 triatomine bugs corresponding to four genera in different departments of Colombia. DNA extraction and PCR were performed to evaluate T. cruzi presence and to determine the discrete typing unit (DTU) of the parasite. PCR products of the bacterial 16S rRNA gene were pooled and sequenced. Resulting reads were denoised and QIIME 2 was used for the identification of amplicon sequence variants (ASVs). Diversity (alpha and beta diversity) and richness analyses, Circos plots, and principal component analysis (PCA) were also performed. The overall T. cruzi infection frequency was 75.9%, with TcI being the predominant DTU. Approximately 500,000 sequences were analyzed and 27 bacterial phyla were identified. The most abundant phyla were Proteobacteria (33.9%), Actinobacteria (32.4%), Firmicutes (19.6%), and Bacteroidetes (7.6%), which together accounted for over 90% of the gut communities identified in this study. Genera were identified for these main bacterial phyla, revealing the presence of important bacteria such as Rhodococcus, Serratia, and Wolbachia. The composition of bacterial phyla in the gut of the insects was significantly different between triatomine species, whereas no significant difference was seen between the state of T. cruzi infection. We suggest further investigation with the evaluation of additional variables and a larger sample size. To our knowledge, this study is the first characterization of the gut bacterial structure of the main triatomine genera in Colombia.
Cardiovascular disease (CVD) is the leading cause of mortality worldwide and in Colombia. The analysis of CVD mortality has been mainly relied on individual factors and rates, but occurrence is also ...related to contextual conditions. Understanding the distribution of CVD in a region will contribute to implement more focused-preventive and care interventions.
Using the national mortality register established by the Department of National Statistics, standardized rates and spatial distribution of CVD mortality were estimated for Cali, Colombia, between 2010-2017. Global and local spatial aggregation was assessed using the Geary's C test and for each district standardized mortality ratios smoothed by the Bayesian empirical were estimated.
Over the period, CVD was the main cause of mortality with 28,804 deaths accounting for 23,8% of total deaths. The global CVD mortality rate varied from 235.9 to 257.4 per 100.000 habitants, with an average increase of 9.1% in the percentage change. The main cause of mortality were hypertensive diseases following by ischemic heart diseases and stroke. The standardized mortality ratios smoothed by the Bayesian empirical method showed that the districts 7, 13, 14, 15 and 16 located at the eastern area with the lowest socio-economic strata and the district 22 at the south of the city with the highest socio-economic strata had the high risks of CVD mortality. All these areas were at the boundary of the city. The the lowest risk was observed in districts 1 and 2 at the northwest area with the upper socio-economic strata. Over the study period, a spatial autocorrelation was found in the districts 1,9 10, 11, 12, 13, 14, 15, 19, and 21 by using the Geary's C test. The highest significant spatial association was found in the districts 1 and 21.
Of 22 districts in Cali, the highest risk of CVD mortality was found in three at the lowest and one in the upper socio-economic strata between 2013 and 2017. Over the period a global spatial aggregation was identified due mainly to districts peripherical located suggesting that there could be contextual conditions influencing the risk. Therefore, there is a need for considering local conditions to prevent CVD mortality.
Triatomines are hematophagous insects that play an important role as vectors of Trypanosoma cruzi, the causative agent of Chagas disease. These insects have adapted to multiple blood-feeding sources ...that can affect relevant aspects of their life-cycle and interactions, thereby influencing parasitic transmission dynamics. We conducted a characterization of the feeding sources of individuals from the primary circulating triatomine genera in Colombia using amplicon-based next-generation sequencing (NGS).
We used 42 triatomines collected in different departments of Colombia. DNA was extracted from the gut. The presence of T. cruzi was identified using real-time PCR, and discrete typing units (DTUs) were determined by conventional PCR. For blood-feeding source identification, PCR products of the vertebrate 12S rRNA gene were obtained and sequenced by next-generation sequencing (NGS). Blood-meal sources were inferred using blastn against a curated reference dataset containing the 12S rRNA sequences belonging to vertebrates with a distribution in South America that represent a potential feeding source for triatomine bugs. Mean and median comparison tests were performed to evaluate differences in triatomine blood-feeding sources, infection state, and geographical regions. Lastly, the inverse Simpson's diversity index was calculated.
The overall frequency of T. cruzi infection was 83.3%. TcI was found as the most predominant DTU (65.7%). A total of 67 feeding sources were detected from the analyses of approximately 7 million reads. The predominant feeding source found was Homo sapiens (76.8%), followed by birds (10.5%), artiodactyls (4.4%), and non-human primates (3.9%). There were differences among numerous feeding sources of triatomines of different species. The diversity of feeding sources also differed depending on the presence of T. cruzi.
To the best of our knowledge, this is the first study to employ amplicon-based NGS of the 12S rRNA gene to depict blood-feeding sources of multiple triatomine species collected in different regions of Colombia. Our findings report a striking read diversity that has not been reported previously. This is a powerful approach to unravel transmission dynamics at microgeographical levels.
Background. Antiretroviral drugs with a lower potential to induce hepatic steatosis in human immunodeficiency virus (HIV) infection need to be identified. We compared the effect of switching ...efavirenz (EFV) to raltegravir (RAL) on hepatic steatosis among HIV-infected patients with nonalcoholic fatty liver disease (NAFLD) receiving EFV plus 2 nucleoside analogues. Methods. HIV-infected patients on EFV plus tenofovir/emtricitabine or abacavir/lamivudine with NAFLD were randomized 1:1 to switch from EFV to RAL (400 mg twice daily), maintaining nucleoside analogues unchanged, or to continue with EFV plus 2 nucleoside analogues. At baseline, eligible patients should show controlled attenuation parameter (CAP) values ≥238 dB/m. Changes in hepatic steatosis at 48 weeks of follow-up over baseline levels were measured by CAP. Results. Overall, 39 patients were included, and 19 of them were randomized to switch to RAL. At week 48, median CAP for the RAL group was 250 (Q1–Q3, 221–277) dB/m and 286 (Q1–Q3, 269–314) dB/m for the EFV group (P = .035). The median decrease in CAP values was −20 (Q1–Q3, −67 to 15) dB/m for the RAL arm and 30 (Q1–Q3, −17 to 49) dB/m for the EFV group (P = .011). CAP values <238 dB/m at week 48 were observed in 9 (47%) patients on RAL and 3 (15%) individuals on EFV (P = .029). Conclusions. After 48 weeks, HIV-infected individuals switching EFV to RAL showed decreases in the degree of hepatic steatosis, as measured by CAP, compared with those continuing with EFV. In addition, the proportion of patients without significant hepatic steatosis after 48 weeks was greater for those who switched to RAL. Clinical Trials Registration. NCT01900015.
We report on the formation of two novel multifunctional isomorphous (4,4) square-grid 2D coordination polymers based on 1H-indazole-5-carboxylic acid. To the best of our knowledge, these complexes ...are the first examples of 2D-coordination polymers constructed with this novel ligand. We have analysed in detail the structural, magnetic and anti-parasitic properties of the resulting materials. In addition, the capability of inhibiting nitric oxide production from macrophage cells has been measured and was used as an indirect measure of the anti-inflammatory response. Finally, the photocatalytic activity was measured with a model pollutant, i.e. vanillic acid (phenolic compound), with the aim of further increasing the functionalities and applicability of the compounds.
We report on the formation of two isomorphous (4,4) square-grid 2D coordination polymers based on 1H-indazole-5-carboxylic acid. These materials show interesting magnetic, anti-parasitic and anti-inflammatory properties. Display omitted
•New 2D coordination polymers based on 1H-indazole-5-carboxylic acid are reported.•Magnetic studies have been performed.•Compounds show interesting anti-parasitic properties.•Anti-inflammatory activities have been analysed.•Cytotoxicity in macrophages has been measured.
This study deals with the extraction, optimization, and evaluation of the antioxidant and antimicrobial activities of bioactive compounds obtained from the seeds of annatto using microwave-assisted ...extraction as compared to leaching. Annatto seeds were subjected to a microwave treatment of 2450 MHz and power of 700 watts using a response surface design involving four factors: pH (4⁻11), solvent concentration (ethanol) (50⁻96 %), solvent-to-seed ratio (2⁻10), and microwave exposure time (0⁻5 min). The contents of polyphenol compounds and bixin were taken as response variables. Subsequently, the antioxidant and antimicrobial activities were assessed at the optimal processing conditions predicted by the experimental design. Microwaves, solvent concentration, and the solvent-to-seed ratio showed a statistically significant effect for the extraction of polyphenol compounds and bixin. Thus, microwaves accelerated the extraction of those compounds and the slight increase in temperature caused some degradation of the polyphenol compounds. The microwave-assisted extraction increased the contents of polyphenols and bixin along with their antioxidant activity as compared to leaching extraction. However, this technique does not significantly improve the antimicrobial activity against
and
.
Background. During peritoneal dialysis (PD), mesothelial cells (MC) undergo an epithelial-to-mesenchymal transition (EMT), and this process is associated with peritoneal membrane (PM) damage. Bone ...morphogenic protein-7 (BMP-7) antagonizes transforming growth factor (TGF)-β1, modulates EMT and protects against fibrosis. Herein, we analysed the modulating role of BMP-7 on EMT of MC in vitro and its protective effects in a rat PD model. Methods. Epitheliod or non-epitheliod MC were analysed for the expression of BMP-7, TGF-β1, activated Smads, epithelial cadherin (E-cadherin), collagen I, alpha smooth muscle cell actin (α-SMA) and vascular endothelial growth factor (VEGF) using standard procedures. Rats were daily instilled with PD fluid with or without BMP-7 during 5 weeks. Histological analyses were carried out in parietal peritoneum. Fibrosis was quantified with van Gieson or Masson's trichrome staining. Vasculature, activated macrophages and invading MC were quantified by immunofluorescence analysis. Quantification of infiltrating leukocytes and MC density in liver imprints was performed by May–Grünwald–Giemsa staining. Hyaluronic acid levels were determined by ELISA. Results. MC constitutively expressed BMP-7, and its expression was downregulated during EMT. Treatment with recombinant BMP-7 resulted in blockade of TGF-β1-induced EMT of MC. We provide evidence of a Smad-dependent mechanism for the blockade of EMT. Exposure of rat peritoneum to PD fluid resulted in inflammatory and regenerative responses, invasion of the compact zone by MC, fibrosis and angiogenesis. Administration of BMP-7 decreased the number of invading MC and reduced fibrosis and angiogenesis. In contrast, BMP-7 had no effect on inflammatory and regenerative responses, suggesting that these are EMT-independent, and probably upstream, processes. Conclusions. Data point to a balance between BMP-7 and TGF-β1 in the control of EMT and indicate that blockade of EMT may be a therapeutic approach to ameliorate peritoneal membrane damage during PD.
Background:
During peritoneal dialysis (PD), the peritoneum is exposed to bioincompatible dialysis fluids that cause epithelial-to-mesenchymal transition of mesothelial cells, fibrosis, and ...angiogenesis. Ultrafiltration failure is associated with high transport rates and increased vascular surface, indicating the implication of vascular endothelial growth factor (VEGF). Sources of VEGF in vivo in PD patients remain unclear. We analyzed the correlation between epithelial-to-mesenchymal transition of mesothelial cells and both VEGF level and peritoneal functional decline.
Methods:
Effluent mesothelial cells were isolated from 37 PD patients and analyzed for mesenchymal conversion. Mass transfer coefficient for creatinine (Cr-MTC) was used to evaluate peritoneal function. VEGF concentration was measured by using standard procedures. Peritoneal biopsy specimens from 12 PD patients and 6 controls were analyzed immunohistochemically for VEGF and cytokeratin expression.
Results:
Nonepithelioid mesothelial cells from effluent produced a greater amount of VEGF ex vivo than epithelial-like mesothelial cells (
P < 0.001). Patients whose drainage contained nonepithelioid mesothelial cells had greater serum VEGF levels than those with epithelial-like mesothelial cells in their effluent (
P < 0.01). VEGF production ex vivo by effluent mesothelial cells correlated with serum VEGF level (
r = 0.6;
P < 0.01). In addition, Cr-MTC correlated with VEGF levels in culture (
r = 0.8;
P < 0.001) and serum (
r = 0.35;
P < 0.05). Cr-MTC also was associated with mesothelial cell phenotype. VEGF expression in stromal cells, retaining mesothelial markers, was observed in peritoneal biopsy specimens from high-transporter patients.
Conclusion:
These results suggest that mesothelial cells that have undergone epithelial-to-mesenchymal transition are the main source of VEGF in PD patients and therefore may be responsible for a high peritoneal transport rate.
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